MBTP 18: Immunophenotyping of NK cells from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 18 (MBTP 18)

This application protocol describes the flow cytometric analysis of NK cells after spleen dissociation from healthy C57BL/6 mice. Mouse spleens are easily dissociated using the gentleMACS™ Dissociators to quickly obtain viable single cell suspensions ready for flow cytometric analysis.

Protocol

Gating strategy showing the analysis of NK cell maturational stages from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described NK cell panel to analyze NK cell maturation stages. Samples were initially gated on single cells using FSC-H/FSC-A gating and on live cells using Viobility-negative gating (data not shown). Mouse NK cells are identified within the CD45+/Lineage (CD3/CD19)-negative population (A) as NK1.1+ and CD335+ cells (B). Mature NK cells express KLRG1 (C). NK cells can further be divided into four maturation stages based upon the expression of CD27 and CD11b (D). Stages in order from less to more mature are: S1<S2<S3<S4.

Materials

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